DETECTION AND QUANTITATION OF ANAPLASMA-MARGINALE IN CARRIER CATTLE BY USING A NUCLEIC-ACID PROBE
DETECTION AND QUANTITATION OF ANAPLASMA-MARGINALE IN CARRIER CATTLE BY USING A NUCLEIC-ACID PROBE
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DOI：
10.1128/jcm.27.2.279-284.1989
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发表时间：
1989-02-01
影响因子：
9.4
通讯作者：
BARBET, AF
中科院分区：
文献类型：
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作者：
ERIKS, IS;PALMER, GH;BARBET, AF
从边缘无形体（一种牛立克次体血寄生虫）急性感染中恢复的牛经常持续感染低水平寄生虫血症，并成为疾病传播的宿主。为了充分了解这些携带者在疾病流行和传播中的作用，必须准确检测和定量低水平的寄生虫血症。我们开发了一种核酸探针，该探针源自编码 105,000 分子量表面蛋白的基因的一部分，可以检测边缘曲霉感染的红细胞。该探针对边缘曲霉具有特异性，可检测0.5毫升血液中0.01纳克的基因组DNA和500至1,000个感染的红细胞，相当于0.000025%的寄生虫血症。这使得探针的灵敏度至少比光学显微镜高 4,000 倍。探针与经处理的来自被证明是无形体病携带者的动物的血液的杂交表明，携带者之间的寄生虫血症水平差异很大，范围从> 0.0025％到<0.000025％。不同日期个体动物的寄生虫血症水平也存在差异。这些结果意味着，在任何给定时间，一群牛内的个体传播疾病的能力可能存在显着差异。
Cattle which have recovered from acute infection with Anaplasma marginale, a rickettsial hemoparasite of cattle, frequently remain persistently infected with a low-level parasitemia and serve as reservoirs for disease transmission. To fully understand the role of these carriers in disease prevalence and transmission, it is essential that low levels of parasitemia can be accurately detected and quantitated. we have developed a nucleic acid probe, derived from a portion of a gene encoding a 105,000-molecular-weight surface protein, that can detect A. marginale-infected erythrocytes. The probe is specific for A. marginale and can detect 0.01 ng of genomic DNA and 500 to 1,000 infected erythrocytes in 0.5 ml of blood, which is equivalent to a parasitemia of 0.000025%. This makes the probe at least 4,000 times more sensitive than light microscopy. Hybridization of the probe with treated blood from animals proven to be carriers of anaplasmosis showed that parasitemia levels were highly variable among carriers, ranging from > 0.0025 to < 0.000025%. Parasitemia levels of individual animals on different dates were also variable. These results imply that, at any given time, individuals within a group of cattle may differ significantly in their abilities to transmit disease.